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Abstract
The full cDNA of an annexin gene from Microcotyle sebastis (MsANX) was cloned for
the first time in monogeneans. The cDNA of MsANX comprises 1199bp with a 29bp 5' untranslated
region, an open reading frame of 1062bp, and a 108bp 3' untranslated region. The recombinantly
produced MsANX bound phosphatidylserine vesicles in the presence of Ca2+, whereas
no MsANX was precipitated in the absence of free Ca2+. Phylogenetically, MsANX formed
a cluster with human annexin A13, known as the earliest annexin in vertebrates and
expressed mainly in the intestine. The localization of MsANX in M. sebastis was analyzed
by Western blotting and immunohistochemistry using the antiserum raised against the
recombinant MsANX. In Western blot analysis, rat antiserum bound to a protein corresponding
to the MsANX in size when worm crude extracts were used as antigens, but no bands
were detected by the antiserum when the excretory/secretory proteins of worms were
used as antigens. In immunohistochemistry analysis, significant antibody binding annexin
was found in the ovarian region, the pharynx and the intestinal caecum of the worm.
Interestingly, the alimentary canal location of MsANX was similar to the location
of human annexin A13, and further research is needed to trace evolutionary relationship
among helminthic annexins and human annexin A13. Also it remains to be investigated
whether immunization of naïve fish with the recombinant MsANX can induce protective
immune responses against M. sebastis infection.